Table 1.
Kinetic characterization of DUBs that are used in Ub chain restriction analysis7 for the diUb FRET pairs 17 a–g.[a]
| DUB | Linkage | K m [μm] | k cat [s−1] | k cat/K m [m −1 s−1] |
|---|---|---|---|---|
| AMSH | Lys63 | 45.4 | 0.0027 | 59 |
| AMSH*[b] | Lys63 | 2.4 | 0.17 | 70 048 |
| Cezanne | Lys11 | 19.4 | 1.5 | 78 818 |
| OTUB1 | Lys48 | ≫50 | n.d. | |
| OTUB1*[b] | Lys48 | 38.6 | 0.66 | 17 158 |
| OTUD1 | Lys63 | ≫50 | n.d. | 4020 |
| OTUD3 | Lys6 | ≫50 | n.d. | |
| Lys11 | 52.4 | 0.0085 | 162 | |
| Lys63 | 57.9 | 0.0061 | 105 | |
| TRABID | Lys29 | 40.1 | 0.053 | 1317 |
| Lys33 | 19.6 | 0.028 | 1431 | |
| Lys63 | 54.0 | 0.034 | 627 | |
| OTUD2 | Lys11 | 87.9 | 4.4 | 50 253 |
| Lys27 | ≫50 | n.d. | ||
| Lys29 | ≫50 | n.d. | ||
| vOTU | Lys6 | 3.6 | 0.87 | 242 031 |
| Lys11 | 3.4 | 0.22 | 63 110 | |
| Lys48 | 8.4 | 1.0 | 119 952 | |
| Lys63 | 1.4 | 0.091 | 66 266 | |
| USP21 | Lys6 | 2.1 | 0.12 | 60 633 |
| Lys11 | 1.4 | 0.087 | 62 735 | |
| Lys33 | 2.1 | 0.13 | 60 814 | |
| Lys48 | 1.7 | 0.067 | 39 181 | |
| Lys63 | 2.7 | 0.16 | 60 558 |
[a] Values in italics were obtained by extrapolation beyond the highest substrate concentration. [b] Activated versions of AMSH and OTUB1 were created by fusing them to their activators (STAM and UBE2D2, respectively).11